Cooray S, Warrener L, Jin L
Virus Reference Department, Centre for Infections, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK.
J Clin Virol. 2006 Jan;35(1):73-80. doi: 10.1016/j.jcv.2004.12.020. Epub 2005 Jul 12.
A reverse transcription nested polymerase chain reaction (RT-PCR) assay was developed and evaluated for detection of rubella virus (RV) RNA directly from clinical specimens using primers that amplified 592 nucleotides, of a variable region within the E1 gene. RV RNA was detected in pre- and post-natal congenital rubella samples and samples from patients with acute rubella, which suggests that it is a reliable technique for rubella diagnosis and surveillance. The sensitivity of the PCR was found to be equivalent to that of previously published assays, which amplify smaller regions of the E1 gene. This improved RT-PCR is much more specific for detection of the rubella genome compared to our previous PCR, where some primers were complementary to the human genome. The larger size of the PCR amplicon was also useful for molecular genotyping of virus strains.
开发并评估了一种逆转录巢式聚合酶链反应（RT-PCR）检测方法，该方法使用能扩增E1基因可变区内592个核苷酸的引物，直接从临床标本中检测风疹病毒（RV）RNA。在先天性风疹的产前和产后样本以及急性风疹患者的样本中检测到了RV RNA，这表明该方法是风疹诊断和监测的可靠技术。发现该PCR的灵敏度与之前发表的扩增E1基因较小区域的检测方法相当。与我们之前的PCR相比，这种改进的RT-PCR在检测风疹基因组方面特异性更强，之前的PCR中一些引物与人基因组互补。PCR扩增子的较大尺寸也有助于病毒株的分子基因分型。
